BIO RAD TC20 User manual

Brand: BIO RAD

Model: TC20

Category: Water pumps

Type: User manual

Catalog #145-0101

TC20

™

Automated Cell Counter

Instruction Manual

without written permission of Bio-Rad Laboratories, Inc. Excel is a trademark of Microsoft Corporation.

TC20

™

Automated

Cell Counter

Instruction Manual

Bio-Rad Laboratories Resources

Table 1 lists Bio-Rad resources and how to locate what you need.

Table 1. Bio-Rad resources.

Resource How to Contact

Local Bio-Rad Laboratories Find information and contacts on the Bio-Rad Laboratories

representatives website by selecting your country on the Home page

(www.bio-rad.com). Find the nearest international office listed on

the back of this manual

Technical notes and literature Go to the Bio-Rad Laboratories website (www.bio-rad.com).

Type a term in the Search box and select Documents to find links

to literature

Technical specialists Bio-Rad Laboratories Technical Support scientists provide our

customers with practical and expert solutions. To find local

Technical Support on the phone, contact your nearest Bio-Rad

Laboratories office. For Technical Support in the United States

and Canada, call 1-800-424-6723 (toll-free phone), and select the

Technical Support option

Warranty

The TC20

™

automated cell counter and associated accessories are covered by a standard

Bio-Rad warranty. Contact your local Bio-Rad Laboratories office for the details of the warranty.

Safety Use Speciﬁcations and Compliance

A Warning! label in this manual warns you about sources of injury or harm, including risk of

electrical shock.

Warning! Do not attempt to repair or remove the outer case of the TC20 automated cell counter

or other accessories. If you open this instrument, you put yourself at risk for harm to body or equipment

from electrical shock.

This instrument has been tested and found to be in compliance with all applicable requirements

of the following safety and electromagnetic standards.

Table 2. Environmental conditions for safe operation of the TC20 automated cell counter.

Transient category Installation categories II

Operating power 90–240 VAC, 1.2 A

Frequency 50/60 Hz

Electrical input 12 VDC, 2.5 A

Installation site Indoor use only

Operating temperature 10–35°C

Maximum relative humidity 20–80%

Altitude <2,400 m

Pollution degree 2

Degree of protection IPX0

Note: Do not store or operate the unit near a sink as contact with water could cause electric shock.

Unit is heavy, do not store or operate it at the edge of a laboratory bench. During operation unit

must be in upright position.

Do not tip the unit over if the counting slide is inserted in the slide port; this could cause accidental

spillage of sample and contaminate the unit.

Use only Bio-Rad counting slides, using other manufacturers´ slides could damage the unit.

TC20 cell counter is factory calibrated, no further calibration is needed.

iii

Safety Compliance

This instrument has been tested and found to be in compliance with all applicable requirements of the

following safety and electromagnetic standards:

• EN61010-1 — Electrical Equipment for Measurement, Control, and Laboratory Use

• UL Std No. 61010A-1 — Electrical Equipment for Measurement, Control, and Laboratory Use, Part

1: General Requirements

• UL Std No. 61010A-2-010 — Electrical Equipment for Measurement, Control, and Laboratory Use,

Part 1: General Requirements

• CAN/CSA C22.2 No. 1010.1-92 — Safety Requirements for Electrical Equipment for Measurement,

Control, and Laboratory Use, Part 1: General Requirements (includes Amendment 1)

• CAN/CSA C22.2 No. 1010.1B-97 Amendment 2 CAN/CSA C22.2 No. 1010.1-92 — Safety

Requirements for Electrical Equipment for Measurement, Control, and Laboratory Use, Part 1:

General Requirements

• IEC 61010-1 — Safety Requirements for Electrical Equipment for Measurement, Control, and

Laboratory Use, Part 1 General Requirements

Electromagnetic Compatibility (EMC)

• F.C.C. Title 47 Part 15B as a Class A digital device

• EN61326 Class A Electrical Equipment for measurement, control, and laboratory use —

General Requirements

FCC Warning and Notes

• Warning: Changes or modifications to this unit not expressly approved by the party responsible for

compliance could void the user’s authority to operate the equipment

• Note: This equipment has been tested and found to comply with the limits for a Class A digital

device, pursuant to part 15 of the FCC Rules. These limits are designed to provide reasonable

protection against harmful interference when the equipment is operated in a commercial

environment. This equipment generates, uses, and can radiate radio frequency energy and, if not

installed and used in accordance with the instruction manual, may cause harmful interference to

radio communications. Operation of this equipment in a residential area is likely to cause harmful

interference, in which case the user will be required to correct the interference at his own expense

• Note regarding FCC compliance: Although this design of instrument has been tested and found

to comply with Part 15, Subpart B, of the FCC Rules for a Class A digital device, please note that

this compliance is voluntary, for the instrument qualifies as an “exempted device” under 47 CFR

15.103(c), in regard to the cited FCC regulations in effect at the time of manufacture

• Note regarding Canadian EMC compliance: Le present appareil numerique n’emet pas de bruits

radioelectrique depassant les limites applicables aux appareils numeriques de classe A prescrites

dans le reglement sur le brouillage radioelectrique edicte par le Ministere des Communications

du Canada

This Bio-Rad instrument is designed and certified to meet EN61010* and the EMC requirements

of EN61326 (for Class A) safety standards. Certified products are safe to use when operated in

accordance with the instruction manual. This instrument should not be modified or altered in any way.

Alteration of this instrument will cause the following results:

• Void the manufacturer’s warranty

• Void the EN61010 safety certification

• Create a potential safety hazard

* EN61010 is an internationally accepted electrical safety standard for laboratory instruments.

Bio-Rad Laboratories is not responsible for any injury or damage caused by the use of this instrument

for purposes other than those for which it is intended, or by modifications of the instrument not

performed by Bio-Rad Laboratories or an authorized agent.

We strongly recommend that you follow the safety specifications listed in this section and throughout

this manual. Use only the supplied power cord in the instrument, making sure to choose the plug

adaptor that corresponds to the electrical outlets in your region.

Table of Contents

Bio-Rad Laboratories Resources ............................................................................ii

Warranty .................................................................................................................ii

Safety Use Specifications and Compliance............................................................iii

Chapter 1 — Introduction ................................................................................... 1

Instrument Overview ......................................................................................... 2

User Interface .................................................................................................... 3

TC20

™

Automated Cell Counter Specifications ................................................. 4

Kits, Reagents, and Accessories....................................................................... 4

Chapter 2 — Setting Up the TC20 Automated Cell Counter ............................ 7

System Components......................................................................................... 8

Unpacking the System Components................................................................. 8

Setting the Date and Time................................................................................. 8

Setting up Gating .............................................................................................. 9

Setting up Automatic Data Export and Sample Naming ................................. 10

Chapter 3 — Performing Cell Counts .............................................................. 11

Loading Slides ................................................................................................. 12

Preparing Samples .......................................................................................... 12

Preparing Samples without Trypan Blue Dye ............................................. 12

Preparing Samples with Trypan Blue Dye .................................................. 12

Counting Cells on the TC20 Automated Cell Counter ..................................... 13

Counting Samples with Multiple Cell Populations ........................................... 14

Sample Naming............................................................................................... 14

Current Count Options .................................................................................... 15

Using the Dilution Calculator ...................................................................... 15

Printing the Current Cell Count .................................................................. 16

Viewing the Image of Cells from the Current Count ................................... 16

Exporting the Image of Cells from the Current Count ................................ 16

Viewing the Histogram and Gating from the Current Count ....................... 17

Chapter 4 — Saved Count Results ................................................................... 19

Viewing Previous Counts ................................................................................. 20

Printing Previous Counts ................................................................................. 20

Exporting Previous Counts .............................................................................. 20

Automatic Image Export.................................................................................. 21

Deleting All Counts .......................................................................................... 22

Chapter 5 — Maintenance and Troubleshooting ............................................ 23

Performing the TC20 System Verification Test ................................................ 24

Cleaning the TC20 Automated Cell Counter.................................................... 25

Thermal Printer Maintenance ..................................................................... 25

Updating the Firmware .................................................................................... 26

Battery ............................................................................................................ 26

Troubleshooting ............................................................................................... 27

Ordering Information ......................................................................................... 28

Chapter 1 — Introduction

Introduction

The Bio-Rad TC20

™

automated cell counter is a benchtop instrument that counts

mammalian cells in one simple step, initiating auto-counts immediately upon slide

insertion. The innovative autofocus technology and sophisticated digital image

analysis algorithm produce accurate cell counts for most samples in less than

30 seconds. The TC20 cell counter rapidly provides a total cell count with or without

trypan blue staining and assesses cell viability via trypan blue exclusion. Getting all

the data you need about your cell cultures is fast and easy; the TC20 cell counter and

disposable counting slides eliminate the need for tedious setup, cleaning, or maintenance.

The TC20 cell counter provides accurate and reproducible cell counts for suspended

cells that have been grown as adherent or suspension cells at concentrations within the

5 × 10

to 1 × 10

cells/ml range and for cells within the 6–50 μm cell diameter range.

Only 10 μl of cell suspension is required for the cell count. The cell counter detects the

presence of trypan blue in the sample with no user input required and automatically

accounts for the 1:1 (trypan blue:cells) dilution factor. The TC20 cell counter can count

single or mixed cell population samples. For samples with multiple cell populations the

user can deﬁne the cell population of interest by adjusting the position of cell size gates.

An image of the counted cells can be viewed onscreen and downloaded onto a USB

ﬂash drive immediately after the count. The TC20 cell counter can store results from

100 counts. Count results can be exported onto a USB ﬂash drive and viewed in

Microsoft Excel. Each sample can be given a unique name and if the data autosave

function is enabled, the JPEG image from the current count is automatically saved onto

the inserted USB ﬂash drive. Each image contains a metatag with the cell count data

and can be further analyzed using the TC20 Data Analyzer downloadable from

www.bio-rad.com/tc20.

The TC20 cell counter has a dilution calculator to derive cell resuspension volumes

required to achieve the cell concentration needed for the next experiment. A histogram

of cell diameter distribution in the sample can be viewed after the cell count.

Instrument Overview

The front panel of the TC20 automated cell counter (Figure 1) includes:

• User interface — a color digital screen and four keys for navigating the

software

• Slide port — used to insert the cell counting slide

The side (right) panel of the TC10 automated cell counter (Figure 1) includes:

• USB port A (top port) — for connecting thermal printer or USB ﬂash drive

• USB port B (bottom port) — for connecting thermal printer; the USB port B

cannot be used for the USB ﬂash drive

• Power switch — for turning the instrument on and off

The rear panel of the TC20 automated cell counter includes:

• Power inlet — used to connect the cell counter to an electrical outlet via the

supplied power cord (select the appropriate plug adaptor for your region)

LCD screen

Keys for navigation

USB ports A and B

Slide port

Power switch

Fig. 1. TC20 cell counter.

Fig. 2. Home screen.

Introduction

User Interface

The user interface contains a color LCD

screen and the four navigation keys

listed below:

• Home — for accessing the

Home screen or for exiting

any screen

• Up and down arrow keys —

for moving to the desired

setting within a screen

• Enter — for conﬁrming a

selection

Home Screen

The Home screen can be accessed from any screen by pressing Home. This screen

(Figure 2) provides access to the following choices:

Count cells — initiates a cell count. When this option is selected, the instrument will

automatically start counting if a slide is already inserted.

If there is no slide in the instrument, the Insert slide screen will be displayed

Current count — for returning to the current count screen

Gating setup — for setting up histogram gating parameters

Previous counts — for viewing results from up to 100 previous counts stored in the

instrument; results are organized by date/time stamps, with the most recent count

shown ﬁrst

Export data — for exporting results and image from the most recent count from the

instrument onto a USB ﬂash drive

Fig. 3. Options screen.

Introduction

Options — for selecting additional operations

Options Screen

The Options screen (Figure 3) contains

the following choices:

Autosave/sample name — for setting

up the automatic export of cell images

to a USB ﬂash drive and for setting up

sample naming preferences

System test — for verifying the

performance of the instrument;

a veriﬁcation slide is needed for

this function

Set date/time — for setting the date

and time on the instrument, results are

associated with date/time stamps

Update ﬁrmware — for upgrading the ﬁrmware

Delete all counts — for deleting all counts stored in the instrument

TC20 Automated Cell Counter Speciﬁcations

Processing time: 30 sec (may increase for higher concentration cell suspensions)

Cell concentration range: 5 × 10

–1 × 10

cells/ml

Optimal concentration range: 1 x 10

–5 x 10

cells/ml

Cell diameter range: 6–50 µm

Dimensions (W x D x H): 19 × 15 × 25.4 cm

Weight: 2.2 kg

Kits, Reagents, and Accessories

TC20 kits, reagents, and accessories are specially designed for use with the

TC20 automated cell counter. These products are optimized to ensure accurate and

reproducible cell counts.

Counting Slides

The counting slide (Figure 4) is a disposable polygon plastic slide with a dual chamber

composed of polymethyl methacrylate (PMMA). Each slide is 75 × 25 × 1.8 mm (W x D x

H) with a chamber depth of 100 μm. Each enclosed chamber requires only 10 μl of

suspended cells. The volume counted in each chamber is 0.4 µl, equivalent to the

volume counted in four squares (each 1 × 1 mm) of a manual hemocytometer.

The slide can provide counts for two different samples or duplicates of the same sample,

one at a time. The chambers are labeled A and B, with an arrow next to each chamber

to indicate the direction of slide insertion. Both slide chambers can be loaded prior to

insertion in the TC20 cell counter for sequential counting.

Fig. 4. Counting slides and trypan blue dye.

Fig. 5. Accessories for the TC20 cell counter.

Introduction

Trypan Blue Dye

The Trypan blue dye has been specially formulated for use with the TC20 counter to

provide rapid staining for accurate determination of cell viability. The dye is composed

of ﬁlter-sterilized 0.4% trypan blue dye (w/v) in 0.81% sodium chloride and 0.06%

potassium phosphate dibasic solution.

Trypan blue is a vital stain recommended for use in estimating the number of

live and dead cells in the cell suspension. The activity of this dye is based on the

fact that the chromophore is negatively charged and does not react with the internal

region of the cell unless the membrane is damaged. The dye enters into only dead

cells and stains them blue, while live cells have intact cell membranes and remain

bright and unstained.

Thermal Label Printer

The thermal label printer (Figure 5) can be connected to the TC20 cell counter via the

USB port A or USB port B. The printer, which uses a direct thermal printing method,

provides labels with results that can be placed into a laboratory notebook.

System Veriﬁcation Kit

The veriﬁcation kit (Figure 5) is available

to verify the instrument’s performance.

The veriﬁcation kit includes a veriﬁcation

slide and instructions for use.

Chapter 2 —

Setting Up the TC20

™

Automated Cell Counter

Fig. 6. Setting the date and time.

Setting Up the TC20 Automated Cell Counter

System Components

The TC20 automated cell counter (catalog #145-0102) includes:

• TC20 automated cell counter

• Power cord with four regional adaptors

• USB ﬂash drive

• Instruction manual

• Quick guide

• Counting slides with dual chambers, 1 box of 30

• Trypan blue dye, 1.5 ml

Accessories that need to be purchased separately include:

• Label printer, 185 labels/roll

• Veriﬁcation kit

Unpacking the System Components

1. Unpack the TC20 automated cell counter carefully. Remove all packaging materials and

store them for future use. Examine the instrument carefully for any damage incurred during

transit. Ensure that all parts of the instrument including accessories listed above are included

with the product. If any item is missing or damaged, contact your local Bio-Rad ofﬁce.

2. Place the TC20 cell counter into upright position on a dry, leveled surface.

3. Insert the supplied power cord into the instrument.

4. Plug the power cord into the appropriate electrical outlet.

5. Turn on the instrument using the green power switch button. The Home

screen appears.

Setting the Date and Time

The TC20 cell counter uses a date/time stamp to track cell count results stored on the

instrument. The date and time should be set on the TC20 cell counter before the ﬁrst count.

Resetting the date/time stamp after using the TC20 cell counter will not affect stored results.

1. Press the down arrow key to go to Options from the Home screen, and press Enter.

2. Use the down arrow key to go to Set

date/time, then press Enter.

3. Use the up or down arrow key to

move through the settings until you

reach the setting you want to modify

(for example, month) (Figure 6). The

press Enter to ﬁnd the correct option

(for example September, October).

4. Use the up or down key to move to

the next setting (for example, day) to

be modiﬁed, and press Enter to ﬁnd

the correct option within the setting.

Setting up the TC20 Automated Cell Counter

5. Finally, when you have made all the modiﬁcations use the up or down arrow key to

reach the Yes or No options for the Save date and time? setting.

6. Press Enter to select Yes to save the changes and return to the Home screen or

select No to return to the Home screen without saving the changes.

Setting up Gating

TC20 cell counter has two counting modes, automated and user deﬁned. In the

automated mode (User deﬁned gates are Disabled) the cell counting algorithm

automatically identiﬁes the cell population of interest. This mode does not require any

user input and is suitable for immortal cell lines and samples that are composed of cells

with similar cell size.

In the user deﬁned mode (User deﬁned gates are Enabled) the cell counter will count only

objects within a user deﬁned cell diameter range, this can be done by adjusting position

of the low and high cell size gate (in

µm). Only objects within the range will be analyzed as

cells, those with diameters outside of that size range will be excluded from the cell count.

Enable user deﬁned gates when counting samples containing multiple cell populations

with a wide range of cell diameters from which you need to select a population of

interest, for example ﬁbroblasts with lymphocyte background.

User deﬁned gates — select Enabled option to switch to the user deﬁned mode or

select Disabled option to switch to the automated mode. Once enabled, a histogram

showing all objects found in the sample will be displayed at the beginning of each

count. Move the size gates to select the range of cell diameters the TC20 cell counter

will include in the count.

Use saved gates — the Yes option will save the position of the size gates from the

previous count for the next count, thus saving time when multiple sample replicates are

counted as you may need to move the size gates on the pre-count histogram only a few

microns or not at all.

If you select No, the size gates will ﬂank what the TC20 cell counter presumes to be the

population of interest.

Image preview — select On to display image of cells with diameters between the

selected cell size gates. No image preview will be displayed if the Off option is selected.

To enable the User deﬁned mode:

1. Press the down arrow key to go to

Gating setup (Figure 7) from the

Home screen, and press Enter.

2. Use the up or down arrow key to

move through the settings (for example,

User deﬁned gates, Image preview)

until you reach the setting you want

to modify. Then press Enter until you

ﬁnd the correct option (for example,

Enabled, Disabled).

3. Use the up or down arrow key to move to

the next setting to be modiﬁed, press Enter

to ﬁnd the correct option for the setting.

Setting up the TC20 Automated Cell Counter

Fig. 7. Setting up gating.

4. Finally, when you have made all the modiﬁcations use the up or down key to reach

Yes or No options for the Save changes? setting.

5. Press Enter to select Yes to save the changes and return to the Home screen, or

select No to return to the Home screen without saving the changes.

Setting Up Automatic Data Export and Sample Naming

In this menu you can enable automatic export of cell image from the most recent

count to a USB ﬂash drive; the image can be viewed on your computer in JPEG format.

You can also use this menu to set sample naming preferences. Enabling sample

name will allow you to select a name consisting of maximum eight alphanumeric

characters. To save time the name selected for the most recent count will automatically

be populated for the next cell count. You can then modify it, for example manually

serialize it. When the automatic name serialization is enabled you can select a

name consisting of maximum six characters, the last two characters are reserved for

automatic numerical serialization. This unique sample name will be used as the JPEG

ﬁle name and will be part of the result record listed in the Previous counts.

Important: At each new cell count the image from previous count is replaced by the

image from the most recent count. When the instrument is switched off, the image

from the last count is no longer kept in memory.

If the Data autosave is disabled, then the Sample name and Serialize sample name

features are automatically disabled.

TC20 cell counter checks sample names already saved on the USB ﬂash drive. If a sample

name is already present on the ﬂash drive, then you will be prompted to select another

name. If a USB ﬂash drive is not inserted, you will be prompted to insert it into USB port A.

To enable the sample naming and automatic data export:

1. Press the down arrow key to go to Options from the Home screen, and press Enter.

2. Press the down arrow key to go to Autosave/sample name (Figure 8), and press Enter.

3. Use the up or down arrow key to move through the settings (for example Data

autosave) until you reach the setting

you want to modify. Then press Enter

until you ﬁnd the correct option (for

example, Enabled, Disabled).

4. Use the up or down arrow key to move

to the next setting to be modiﬁed, press

Enter to ﬁnd the correct option for the

setting.

5. Finally, when you have made all the

modiﬁcations use the up or down key

to reach Yes or No options for the

Save changes? setting.

6. Press Enter to select Yes to save

the changes and return to the Home

screen, or select No to return to the

Home screen without saving the changes.

Setting up the TC20 Automated Cell Counter

Fig. 8. Setting up data export and sample naming.

Chapter 3 —

Performing Cell Counts

Fig. 9. Loading the counting slide.

Performing Cell Counts

Loading Slides

Handle the TC20

™

counting slides using the

edges and avoid touching the optical surface of

the slides.

Important: When loading the sample, place

the pipet tip at a 45° angle at the bottom of the

sample loading area (half circle at outer end of

the chambers). Slide the tip along the surface

and carefully touch the apex of the half circle

(Figure 9). Once the tip is stopped, depress the

plunger to begin the capillary loading process.

Care should be taken to avoid visible bubble

formation or back splatter. Do not overﬁll or

underﬁll the chamber. Overﬁlling the chamber and possible resulting accidental

spillage of sample inside the instrument could lead to biological contamination of the

cell counter.

The cell counting slides cannot be reused. Dispose of used slides as biohazardous waste

according to your local environmental health and safety regulations. To avoid injury do not

break the counting slides.

Preparing Samples

Preparing Samples without Trypan Blue Dye

1. Pipet 10 μl of the cell suspension into the outer opening of either chamber of the

counting slide (Figure 9).

Preparing Samples with Trypan Blue

1. To determine cell viability, mix 1 part trypan blue dye and 1 part cell suspension: In

a micro test tube or on Paraﬁlm combine 10 μl of the cell suspension with 10 μl of

trypan blue dye. Gently pipet up and down ten times to mix.

2. Pipet 10 μl of the mixture into the opening of either chamber on the counting slide.

3. When counting the sample in duplicate, combine 20 μl of the cell suspension with

20 μl of trypan blue dye, and then pipet 10 μl of the mixture into each chamber.

Important: The cell suspension must be loaded into the counting slides and counted

immediately (within 5 minutes of mixing with trypan blue dye). Viable cells that are

exposed to trypan blue dye for an extended period may start incorporating the dye,

affecting the accuracy of the cell count.

Make sure the stock cell suspension is thoroughly mixed by pipetting or vortexing.

Failure to do so can result in improper sample representation within the loaded

counting chamber. Pipet the sample from the middle of the tube ﬁlled with stock cell

suspension. Pipetting from the bottom or top can result in a sample with a higher or

lower concentration, respectively.

If you work with adherent cells, use trypsin to get them into suspension. Using trypsin

instead of scraping improves cell roundness and decreases the number of cell clusters.

Fig. 10. Inserting the counting slide into

the TC20 cell counter.

Fig. 11. Cell count screen without

trypan blue dye.

Fig. 12. Cell count screen with trypan

blue dye.

Performing Cell Counts

Counting Cells on the

TC20 Automated Cell Counter

1. Simply inserting a counting slide into the slide

slot of the TC20 cell counter will automatically

initiate a cell count (Figure 10). The chambers

are labeled A and B, with an arrow next to

each chamber to indicate the direction of

slide insertion. Make sure that the slide is

completely inside the slide slot; otherwise

the instrument does not detect the slide and

counting does not start. Make sure the slide

is inserted right side up.

2. The cell counter automatically detects the

presence of the slide and initiates the count.

The cell counter automatically detects the

presence of trypan blue dye.

3. The count results appear on the

Current count screen.

For samples without trypan blue dye —

On the Current count screen, the instrument

provides the total cell count per ml (Figure 11).

For samples with trypan blue dye — On the

Current count screen, the instrument provides

the total cell count per ml, live cell count per

ml, and percentage of live cells (Figure 12). If

the counted sample contains trypan blue, the

instrument accounts for 1:1 dilution of trypan

blue to cell suspension.

Important: If the number of cells is above

or below the speciﬁed range of the TC20

automated cell counter, “Value out of range” is

displayed on the Current count screen.

View the image of the cells (see Viewing the

Image of Cells from the Current Count for

details) and determine whether the sample

should be diluted or concentrated. Then repeat

the cell count.

If the result is between 5 x 10

and 5 x 10

cells/ ml, counting multiple sample replicates will

increase accuracy. After performing replicate measurements calculate the average and use

it as your result. If the result is above 5 x 10

cells/ml, only one measurement is needed.

4. Once the instrument completes the cell count, remove the slide from the slide slot.

To exit the Current count screen, press Home.

5. If the slide is removed, results from the last count will remain on the screen until a new slide

is inserted. When a new slide is inserted and the instrument starts counting, the previous

result will be stored on the TC20 cell counter using a date/time stamp to track the count.

Counting Samples with Multiple Cell Populations

When counting samples with multiple

cell populations that vary greatly in size

(for example, ﬁbroblasts with lymphocyte

background) enable the User deﬁned

gates (Figure 7). A histogram (Figure 13)

will be displayed at the beginning of the

cell count; you can adjust the size gates

to select the population of interest.

Important: This histogram displayed

at the beginning of the count shows

all objects present in the sample. After

you have conﬁrmed cell size range to

be analyzed (delineated by position of

the cell size gates) objects within that

speciﬁed cell size range are analyzed

as cells, those with diameters outside of

that size range are excluded from the cell count.

If User deﬁned gates are Disabled, the TC20 cell counter will automatically (without

input from the user) determine what the population of interest is.

If unsure which part of the histogram represents your population of interest, enable the

manual mode and move the size gates to the extremes of the histogram. Then open the

exported JPEG on your computer using the TC20 data analyzer and move the size gates

until only the cells you are interested in are annotated in the image. Use the displayed size

gate positions (in microns) as a guide for subsequent counts on the TC20 cell counter.

To select a population on the histogram displayed at the beginning of the cell count:

1. When the histogram is displayed the low size gate is ﬂashing and positions of gates

(in microns) are displayed in the lower right corner.

2. Use the up or down arrow keys to move the low gate to the desired position.

3. Press Enter to select the high gate. Use the up or down arrow key to move the high

gate to the desired position.

4. Press Enter to conﬁrm position of both gates.

5. Press Enter to proceed with the count, or press Home to start over. Press Home

anytime during the process to start over.

6. Press Home twice to return to the Home screen.

Sample Naming

When sample naming is enabled (see Setting Up Automatic Data Export and

Sample Naming for details) you will be prompted to select a sample name for each

cell count. You can select a name containing up to eight alphanumeric characters.

This unique sample name will be used as the JPEG ﬁle name and will be part of the

result record listed in the Previous counts.

To save time, the sample name you selected for the previous count will automatically

be populated for the next count and can be modiﬁed, for example manually serialized.

Performing Cell Counts

Fig. 13. Counting samples with multiple cell

populations.

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BIO RAD TC20 User manual

Brand: BIO RAD

Model: TC20

Category: Water pumps

Type: User manual

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